Nuclear SSU rRNA Gene Research Articles

Taxonomic contributions to Hapalidiales (Corallinophycidae, Rhodophyta): Boreolithothamnion gen. nov., Lithothamnion redefined and with three new species and Roseolithon with new combinations.

Phylogenetic analyses of rbcL gene sequences and of concatenated rbcL, psbA, and nuclear SSU rRNA gene sequences resolved the generitype of Lithothamnion, L. muelleri, in a clade with three other southern Australian species, L. kraftii sp. nov., L. saundersii sp. nov., and L. woelkerlingii sp. nov. Cold water boreal species currently classified in Lithothamnion and whose type specimens have been sequenced are transferred to Boreolithothamnion gen. nov., with B. glaciale comb. nov. as the generitype. The other species are B. giganteum comb. nov., B. phymatodeum comb. nov., and B. sonderi comb. nov., whose type specimens are newly sequenced, and B. lemoineae comb. nov., B. soriferum comb. nov., and B. tophiforme comb. nov., whose type specimens were already sequenced. Based on rbcL sequences from the type specimens of Lithothamnion crispatum, L. indicum, and L. superpositum, each is recognized as a distinct species and transferred to the recently described Roseolithon as R. crispatum comb. nov., R. indicum comb. nov., and R. superpositum com. nov., respectively. To correctly assign species to these three genera based only on morpho-anatomy, specimens must have multiporate conceptacles and some epithallial cells with flared walls. The discussion provides examples demonstrating that only with phylogenetic analyses of DNA sequences can the evolution of morpho-anatomical characters of non-geniculate corallines be understood and applied at the correct taxonomic rank. Finally, phylogenetic analyses of DNA sequences support recognition of the Hapalidiales as a distinct order characterized by having multiporate tetra/bisporangial conceptacles, and not as a suborder of Corallinales whose tetra/bisporangial conceptacles are uniporate.

Analysis of autapomorphic point mutations provides a key for the tangled taxonomic distinction of the closely related species, Euplotes crassus, E. minuta and E. vannus (Ciliophora, Euplotida)

A well-defined clade of the Euplotes phylogenetic tree is represented by marine species characterized by a single-type dargyrome and ten fronto-ventral cirri. Three of them, namely Euplotes crassus, E. minuta and E. vannus, form a complex of closely related species of large use in experimental ciliatology. Despite morphometric and genetic analyses having substantiated their taxonomic separation, ambiguities still persist in strain assignments to one or another species. In addition to objective reasons intrinsic to significant overlapping of most morphological parameters, ambiguities also result from divergences (inherited from past literature) in deciding which of the two morphotypes, E. crassus or E. vannus, is characterized by a larger or a medium cell body size (E. minuta being clearly distinct by a smaller morphotype). By analysing nuclear SSU-rRNA gene and ITS region sequences from 37 strains, previously assigned to E. crassus, E. minuta and E. vannus based on conventional taxonomic parameters, we identified and used ITS autapomorphic point mutations to design three species-specific primers. In combination with an Euplotes-generic primer, they proved to be very effective in running polymerase chain reactions that produce amplicons of species-specific size that reliably resolve ambiguities in assigning strains to E. crassus, E. minuta or E. vannus.

Cyphoderia ampulla (Cyphoderiidae: Rhizaria), a tale of freshwater sailors: The causes and consequences of ecological transitions through the salinity barrier in a family of benthic protists.

The salinity barrier that separates marine and freshwater biomes is probably the most important division in biodiversity on Earth. Those organisms that successfully performed this transition had access to new ecosystems while undergoing changes in selective pressure, which often led to major shifts in diversification rates. While these transitions have been extensively investigated in animals, the tempo, mode, and outcome of crossing the salinity barrier have been scarcely studied in other eukaryotes. Here, we reconstructed the evolutionary history of the species complex Cyphoderia ampulla (Euglyphida: Cercozoa: Rhizaria) based on DNA sequences from the nuclear SSU rRNA gene and the mitochondrial cytochrome oxidase subunit I gene, obtained from publicly available environmental DNA data (GeneBank, EukBank) and isolated organisms. A tree calibrated with euglyphid fossils showed that four independent transitions towards freshwater systems occurred from the mid‐Miocene onwards, coincident with important fluctuations in sea level. Ancestral trait reconstructions indicated that the whole family Cyphoderiidae had a marine origin and suggest that ancestors of the freshwater forms were euryhaline and lived in environments with fluctuating salinity. Diversification rates did not show any obvious increase concomitant with ecological transitions, but morphometric analyses indicated that species increased in size and homogenized their morphology after colonizing the new environments. This suggests adaptation to changes in selective pressure exerted by life in freshwater sediments.

Ribovore: ribosomal RNA sequence analysis for GenBank submissions and database curation

BackgroundThe DNA sequences encoding ribosomal RNA genes (rRNAs) are commonly used as markers to identify species, including in metagenomics samples that may combine many organismal communities. The 16S small subunit ribosomal RNA (SSU rRNA) gene is typically used to identify bacterial and archaeal species. The nuclear 18S SSU rRNA gene, and 28S large subunit (LSU) rRNA gene have been used as DNA barcodes and for phylogenetic studies in different eukaryote taxonomic groups. Because of their popularity, the National Center for Biotechnology Information (NCBI) receives a disproportionate number of rRNA sequence submissions and BLAST queries. These sequences vary in quality, length, origin (nuclear, mitochondria, plastid), and organism source and can represent any region of the ribosomal cistron.ResultsTo improve the timely verification of quality, origin and loci boundaries, we developed Ribovore, a software package for sequence analysis of rRNA sequences. The ribotyper and ribosensor programs are used to validate incoming sequences of bacterial and archaeal SSU rRNA. The ribodbmaker program is used to create high-quality datasets of rRNAs from different taxonomic groups. Key algorithmic steps include comparing candidate sequences against rRNA sequence profile hidden Markov models (HMMs) and covariance models of rRNA sequence and secondary-structure conservation, as well as other tests. Nine freely available blastn rRNA databases created and maintained with Ribovore are used for checking incoming GenBank submissions and used by the blastn browser interface at NCBI. Since 2018, Ribovore has been used to analyze more than 50 million prokaryotic SSU rRNA sequences submitted to GenBank, and to select at least 10,435 fungal rRNA RefSeq records from type material of 8350 taxa.ConclusionRibovore combines single-sequence and profile-based methods to improve GenBank processing and analysis of rRNA sequences. It is a standalone, portable, and extensible software package for the alignment, classification and validation of rRNA sequences. Researchers planning on submitting SSU rRNA sequences to GenBank are encouraged to download and use Ribovore to analyze their sequences prior to submission to determine which sequences are likely to be automatically accepted into GenBank.

Ubiquity of Euglena mutabilis Population in Three Ecologically Distinct Acidic Habitats in Southwestern Japan

Three strains of Euglena mutabilis were isolated from sediments in acidic inland water systems (pH = 3.4–4.7), in Southwestern Japan—acid mine drainage in Sensui (Fukuoka), cold sulfidic spring in Bougatsuru (Oita), and a temporal pool in the Ebinokogen volcanic area (Miyazaki). All strains grew well in acidic media at pH 3.07. Phylogenetic analysis among these three strains showed high similarities to plastid SSU and nuclear SSU rRNA gene sequences (99.86% and 99.76%, respectively). They were closely related to the cultured isolates from other highly acidic habitats (pH = 2.0–5.9). Concentration of sulfate, aluminum, calcium, and iron had 7–70 fold of differences among the three studied habitats. Our results imply that the rRNA genes of E. mutabilis have compensated for their low genetic diversity by adapting to a wide pH range, as well as various water chemistry of habitats.

Molecular and morphological reappraisal of Spyridiocolax capixabus (Spyridiaceae, Rhodophyta), a rare endemic parasite from Brazil

Abstract The monotypic red algal parasite genus Spyridiocolax was proposed from material from the coast of Espírito Santo to accommodate Spyridiocolax capixabus, endemic from Brazil. The species is rare, and we performed a morphological reappraisal and the first molecular characterization of S. capixabus. Parasite specimens growing on Spyridia clavata were collected in the type locality, Itaoca Beach, Espírito Santo. Spyridiocolax capixabus forms colorless warts on the branches of S. clavata, and the secondary pit connections were described for the first time. Male, female and tetrasporophytic structures were observed growing in the same host plant. The plastid rbcL sequences of parasite and host were identical. A divergence of one nucleotide was found to the nuclear SSU rRNA gene, suggesting that the parasite retains the chloroplast of its host. Both plastidial and nuclear phylogenies supported the close relationship of S. capixabus and S. clavata. The data obtained corroborate other studies with rhodophycean parasites, which show morphological and molecular similarities between parasites and hosts. To maintain the monophyly of the host genus, the transfer of S. capixabus to Spyridia is proposed here on the basis of morphological and molecular evidence. Our study constitutes a starting point for reinvestigating the red algal parasites in Brazil.

Detection and quantification of a keystone pathogen in a coastal marine ecosystem

MEPS Marine Ecology Progress Series Contact the journal Facebook Twitter RSS Mailing List Subscribe to our mailing list via Mailchimp HomeLatest VolumeAbout the JournalEditorsTheme Sections MEPS 606:79-90 (2018) - DOI: https://doi.org/10.3354/meps12776 Detection and quantification of a keystone pathogen in a coastal marine ecosystem Robyn Buchwald, Robert E. Scheibling*, Alastair G. B. Simpson Department of Biology, Dalhousie University, Halifax, NS, B3H 4R2, Canada *Corresponding author: rescheib@dal.ca ABSTRACT: The pathogenic amoeba Paramoeba invadens causes recurrent mass mortalities of green sea urchins Strongylocentrotus droebachiensis in coastal Nova Scotia, Canada, driving regime shifts from urchin barrens to kelp beds. Outbreaks of the disease (paramoebiasis) are sporadic, and the source population(s) and epizootiology of the amoeba are poorly understood. We developed PCR-based detection of P. invadens in urchin tissue, sediment, and seawater. Primers specific to the P. invadens nuclear SSU rRNA gene were designed and used in PCR and qPCR analyses to better detect and quantify P. invadens during, following, and in the absence of a natural disease outbreak. A comparison of pathogen load in asymptomatic and symptomatic sea urchins indicated a lower threshold of ~1 cell mg-1 tissue for observing overt signs of paramoebiasis in urchins. P. invadens was detected for the first time in sediment during and following an outbreak of disease in 2014. It also was detected in low abundance (<10 cells l-1) in seawater in fall 2015 in the absence of sea urchin mass mortality or a strong storm event, but not under similar conditions in summer/fall 2016 and 2017. The ability to detect and quantify this pathogen in sea urchins and environmental samples sheds new light on mechanisms of introduction, spread, and persistence of P. invadens along the Nova Scotian coast and the role of large-scale meteorological events and ocean warming in these processes. KEY WORDS: Epizootic · Ecosystem dynamics · Paramoeba invadens · Strongylocentrotus droebachiensis · Quantitative real-time PCR · Pathogenic amoebae · Sea urchins Full text in pdf format Supplementary material PreviousNextCite this article as: Buchwald R, Scheibling RE, Simpson AGB (2018) Detection and quantification of a keystone pathogen in a coastal marine ecosystem. Mar Ecol Prog Ser 606:79-90. https://doi.org/10.3354/meps12776 Export citation RSS - Facebook - Tweet - linkedIn Cited by Published in MEPS Vol. 606. Online publication date: November 15, 2018 Print ISSN: 0171-8630; Online ISSN: 1616-1599 Copyright © 2018 Inter-Research.

Phylogeny of the egg-loving green alga Oophila amblystomatis (Chlamydomonadales) and its response to the herbicides atrazine and 2,4-D

The spotted salamander (Ambystoma maculatum) shares a unique endosymbiotic relationship with the unicellular green alga, Oophila amblystomatis. Despite studies isolating and identifying O. amblystomatis in salamander eggs, the taxonomic identity of the alga remains a point of ongoing debate. In this study, the nuclear SSU rRNA gene was used to characterize two well-supported Oophila clades that include lineages identified from past studies in addition to new isolates from the current study. These two clades do not form a monophyletic group and, furthermore, O. amblystomatis appears to be paraphyletic with numerous other chlamydomonad algae. To gain further insight into the biogeographic variation of the host A. maculatum, the mitochondrial ND4 and control gene regions were examined and the phylogeography was observed to be similar to that noted in the literature. Additionally, the response of O. amblystomatis to atrazine and 2,4-dichlorophenoxyacetic acid following 96 h exposure and a 96 h recovery phase was characterized, as this is a plausible mechanism by which the development of the host salamander could be impaired by herbicides. At the 96 h growth rate, no-observed-effect concentrations were 64 μg/L and 30 mg/L for atrazine and 2,4-D, respectively. We observed full recovery ofO. amblystomatis at these concentrations within 96 h. These data suggest that atrazine and 2,4-D do not pose a significant risk to the symbiotic algae, or, indirectly, to the host salamander. In conclusion, we recommend a revision of the current taxonomy of O. amblystomatis, and demonstrate the need for species identification and thorough phylogenetic reconstruction in toxicity testing to accurately inform risk assessment.

Utility of mitochondrial CO1 sequences for species discrimination of Spirotrichea ciliates (Protozoa, Ciliophora)

Ciliates are a diverse species group of the Protozoa, and nuclear and mitochondrial genes have been utilized to discover new species and discriminate closely related species. The mitochondrial cytochrome c oxidase subunit 1 (CO1) gene has been used to discriminate metazoan species and has also been applied for some groups in the phylum Ciliophora. However, it is difficult to produce a universal primer as a standard barcode, because unlike metazoans, mitochondrial DNA sequences of ciliates are long and highly variable. Therefore, to design the new primer set, we sequenced the mitochondrial genomes of two pseudokeronopsids in the class Spirotrichea using next-generation sequencing technology (HiSeq™ 2000). Based on putative CO1 gene fragments of the pseudokeronopsids, we designed the new primer set and successfully sequenced the CO1 of 69 populations representing 47 species (five orders, 14 families, and 27 genera). We found that CO1 showed higher resolution for separating congeneric species than did nuclear SSU rRNA gene sequences, and we identified some putative cryptic species.

Morphology and molecular phylogeny of Holostichides terrae nov. spec. (Ciliophora: Spirotrichea) with discussion on the possible non-monophyly of Holostichides

In a study to investigate ciliate diversity, we discovered a new soil ciliate. Holostichides terrae nov. spec. was examined and identified based on observations of living cells and stained specimens. In addition, the nuclear SSU rRNA gene along with morphology was analyzed to infer its phylogenetic position. The new species closely resembles H. dumonti, but can be distinguished by the morphology of the pharynx (with rod-shaped structure vs. lacking) and the number of frontoterminal cirri (invariably two vs. usually more than two). Molecular analyses indicate that the genus Holostichides is not monophyletic, and H. terrae is closely related with the genera Birojimia and Hemicycliostyla, both of which have a pharynx with rod-shaped structures, as also seen in H. terrae.

Morphology and Molecular Phylogeny of Two New Brackish Water Ciliates of Bakuella (Ciliophora: Urostylida: Bakuellidae) from South Korea

Two new species of Bakuella, B. (B.) incheonensis n. sp. and B. (Pseudobakuella) litoralis n. sp., were discovered in brackish water near Aamdo Shore Park, Incheon, South Korea. We conducted a morphological study based on live observations and protargol-impregnated specimens, and a molecular analysis using nuclear SSU rRNA gene sequences. Bakuella (B.) incheonensis is diagnosed by: body size 70-105 × 20-40 μm in vivo, 21-25 adoral membranelles, three or four frontoterminal cirri, midventral complex composed of 7-10 midventral pairs with one or two rows and terminating at about 62% of body length, 20-28 left and 25-32 right marginal cirri, 58-87 macronuclear nodules, and yellowish cortical granules. Bakuella (Pseudobakuella) litoralis is diagnosed by: body size 90-125 × 30-40 μm in vivo, 25-33 adoral membranelles, 3-5 buccal cirri, midventral complex composed 10-15 midventral pairs with one or two rows and terminated at 70% of body length, one or two pretransverse cirri, 3-6 transverse cirri, 26-39 left and 29-47 right marginal cirri, 49-84 macronuclear nodules, and two types of cortical granules. Molecular phylogeny using SSU rRNA gene sequences shows a nonmonophyletic relationship among Bakuella species and emphasizes the need for further morphogenetic studies of this genus and other related hypotrichs.

“Cryptic” group-I introns in the nuclear SSU-rRNA gene of Verticillium dahliae

Group-I introns are widespread--though irregularly distributed--in eukaryotic organisms, and they have been extensively used for discrimination and phylogenetic analyses. Within the Verticillium genus, which comprises important phytopathogenic fungi, a group-I intron was previously identified in the SSU-rRNA (18S) gene of only V. longisporum. In this work, we aimed at elucidating the SSU-located intron distribution in V. dahliae and other Verticillium species, and the assessment of heterogeneity regarding intron content among rDNA repeats of fungal strains. Using conserved PCR primers for the amplification of the SSU gene, a structurally similar novel intron (sub-group IC1) was detected in only a few V. dahliae isolates. However, when intron-specific primers were used for the screening of a diverse collection of Verticillium isolates that originally failed to produce intron-containing SSU amplicons, most were found to contain one or both intron types, at variable rDNA repeat numbers. This marked heterogeneity was confirmed with qRT-PCR by testing rDNA copy numbers (varying from 39 to 70 copies per haploid genome) and intron copy ratios in selected isolates. Our results demonstrate that (a) IC1 group-I introns are not specific to V. longisporum within the Verticillium genus, (b) V. dahliae isolates of vegetative compatibility groups (VCGs) 4A and 6, which bear the novel intron at most of their rDNA repeats, are closely related, and (c) there is considerable intra-genomic heterogeneity for the presence or absence of introns among the ribosomal repeats. These findings underline that distributions of introns in the highly heterogeneous repetitive rDNA complex should always be verified with sensitive methods to avoid misleading conclusions for the phylogeny of fungi and other organisms.

Global sampling of plant roots expands the described molecular diversity of arbuscular mycorrhizal fungi

We aimed to enhance understanding of the molecular diversity of arbuscular mycorrhizal fungi (AMF) by building a new global dataset targeting previously unstudied geographical areas. In total, we sampled 96 plant species from 25 sites that encompassed all continents except Antarctica. AMF in plant roots were detected by sequencing the nuclear SSU rRNA gene fragment using either cloning followed by Sanger sequencing or 454-sequencing. A total of 204 AMF phylogroups (virtual taxa, VT) were recorded, increasing the described number of Glomeromycota VT from 308 to 341 globally. Novel VT were detected from 21 sites; three novel but nevertheless widespread VT (Glomus spp. MO-G52, MO-G53, MO-G57) were recorded from six continents. The largest increases in regional VT number were recorded in previously little-studied Oceania and in the boreal and polar climatic zones - this study providing the first molecular data from the latter. Ordination revealed differences in AM fungal communities between different continents and climatic zones, suggesting that both biogeographic history and environmental conditions underlie the global variation of those communities. Our results show that a considerable proportion of Glomeromycota diversity has been recorded in many regions, though further large increases in richness can be expected in remaining unstudied areas.

PHYMYCO-DB: a curated database for analyses of fungal diversity and evolution.

BackgroundIn environmental sequencing studies, fungi can be identified based on nucleic acid sequences, using either highly variable sequences as species barcodes or conserved sequences containing a high-quality phylogenetic signal. For the latter, identification relies on phylogenetic analyses and the adoption of the phylogenetic species concept.Such analysis requires that the reference sequences are well identified and deposited in public-access databases. However, many entries in the public sequence databases are problematic in terms of quality and reliability and these data require screening to ensure correct phylogenetic interpretation.Methods and Principal FindingsTo facilitate phylogenetic inferences and phylogenetic assignment, we introduce a fungal sequence database. The database PHYMYCO-DB comprises fungal sequences from GenBank that have been filtered to satisfy stringent sequence quality criteria. For the first release, two widely used molecular taxonomic markers were chosen: the nuclear SSU rRNA and EF1-α gene sequences. Following the automatic extraction and filtration, a manual curation is performed to remove problematic sequences while preserving relevant sequences useful for phylogenetic studies. As a result of curation, ∼20% of the automatically filtered sequences have been removed from the database. To demonstrate how PHYMYCO-DB can be employed, we test a set of environmental Chytridiomycota sequences obtained from deep sea samples.ConclusionPHYMYCO-DB offers the tools necessary to: (i) extract high quality fungal sequences for each of the 5 fungal phyla, at all taxonomic levels, (ii) extract already performed alignments, to act as ‘reference alignments’, (iii) launch alignments of personal sequences along with stored data. A total of 9120 SSU rRNA and 672 EF1-α high-quality fungal sequences are now available.The PHYMYCO-DB is accessible through the URL http://phymycodb.genouest.org/.

Discovery of New Hydrothermal Activity and Chemosynthetic Fauna on the Central Indian Ridge at 18°–20°S

Indian Ocean hydrothermal vents are believed to represent a novel biogeographic province, and are host to many novel genera and families of animals, potentially indigenous to Indian Ocean hydrothermal systems. In particular, since its discovery in 2001, much attention has been paid to a so-called ‘scaly-foot’ gastropod because of its unique iron-sulfide-coated dermal sclerites and the chemosynthetic symbioses in its various tissues. Despite increasing interest in the faunal assemblages at Indian Ocean hydrothermal vents, only two hydrothermal vent fields have been investigated in the Indian Ocean. Here we report two newly discovered hydrothermal vent fields, the Dodo and Solitaire fields, which are located in the Central Indian Ridge (CIR) segments 16 and 15, respectively. Chemosynthetic faunal communities at the Dodo field are emaciated in size and composition. In contrast, at the Solitaire field, we observed faunal communities that potentially contained almost all genera found at CIR hydrothermal environments to date, and even identified previously unreported taxa. Moreover, a new morphotype of ‘scaly-foot’ gastropod has been found at the Solitaire field. The newly discovered ‘scaly-foot’ gastropod has similar morphological and anatomical features to the previously reported type that inhabits the Kairei field, and both types of ‘scaly-foot’ gastropods genetically belong to the same species according to analyses of their COI gene and nuclear SSU rRNA gene sequences. However, the new morphotype completely lacks an iron-sulfide coating on the sclerites, which had been believed to be a novel feature restricted to ‘scaly-foot’ gastropods. Our new findings at the two newly discovered hydrothermal vent sites provide important insights into the biodiversity and biogeography of vent-endemic ecosystems in the Indian Ocean.

A Second Species ofRhodachlya(Rhodachlyales, Rhodophyta) from Hawaii, with a Description ofR.hawaiianasp. nov.

Rhodachlya hawaiiana Kurihara, J.A. West, Conklin et A.R. Sherwood sp. nov. is described based on cultured material from the island of Hawaii as the second member of the genus Rhodachlya. The pit plug ultrastructure of this alga is characterized by the presence of inner and outer cap layers but lacking a cap membrane between the layers, which is a diagnostic feature of the order Rhodachlyales and the genus Rhodachlya. Morphologically, R. hawaiiana is distinctive from R. madagascarensis in vegetative cell dimensions, frequency of colorless hair cell formation, and spore germination pattern. Rhodachlya hawaiiana and R. madagascarensis are also distinguishable by their nuclear SSU rRNA gene sequence divergence (p-distance = 0.0064), which is comparable to other species-level divergences among members of the Nemaliophycidae.Rhodachlya hawaiiana Kurihara, J.A. West, Conklin et Sherwood sp. nov., décrite à partir de materiel cultivé provenant de l'île d'Hawaii, est la seconde espèce appartenant au genre Rhodachlya. L'ultrastructure des synapses de cette algue montre la présence d'une couche interne et d'une couche externe recouvrant le bouchon synaptique, sans membrane de revêtement entre les couches, ce qui est une des caractéristiques de l'ordre des Rhodachlyales. Morphologiquement R. hawaiiana diffère de R. madagascarensis par la taille de ses cellules végétatives, par la fréquence des cellules formant des poils hyalins et par le mode de germination des spores. Ils diffèrent aussi par la séquence de leur SSU rRNA avec une divergence (p-distance=0.0064) comparable à d'autres divergences entre espèces parmi les Nemaliophycidae.

A NEW LOOK AT AN ANCIENT ORDER: GENERIC REVISION OF THE BANGIALES (RHODOPHYTA)(1).

The red algal order Bangiales has been revised as a result of detailed regional studies and the development of expert local knowledge of Bangiales floras, followed by collaborative global analyses based on wide taxon sampling and molecular analyses. Combined analyses of the nuclear SSU rRNA gene and the plastid RUBISCO LSU (rbcL) gene for 157 Bangiales taxa have been conducted. Fifteen genera of Bangiales, seven filamentous and eight foliose, are recognized. This classification includes five newly described and two resurrected genera. This revision constitutes a major change in understanding relationships and evolution in this order. The genus Porphyra is now restricted to five described species and a number of undescribed species. Other foliose taxa previously placed in Porphyra are now recognized to belong to the genera Boreophyllum gen. nov., Clymene gen. nov., Fuscifolium gen. nov., Lysithea gen. nov., Miuraea gen. nov., Pyropia, and Wildemania. Four of the seven filamentous genera recognized in our analyses already have generic names (Bangia, Dione, Minerva, and Pseudobangia), and are all currently monotypic. The unnamed filamentous genera are clearly composed of multiple species, and few of these species have names. Further research is required: the genus to which the marine taxon Bangia fuscopurpurea belongs is not known, and there are also a large number of species previously described as Porphyra for which nuclear SSU ribosomal RNA (nrSSU) or rbcL sequence data should be obtained so that they can be assigned to the appropriate genus.

PHYLOGENETIC PLACEMENT, GENOME SIZE, AND GC CONTENT OF THE LIQUID‐HYDROCARBON‐PRODUCING GREEN MICROALGA BOTRYOCOCCUS BRAUNII STRAIN BERKELEY (SHOWA) (CHLOROPHYTA)1

We report the genome size and the GC content, and perform a phylogenetic analysis on Botryococcus braunii Kütz., a green, colony‐forming, hydrocarbon‐rich alga that is an attractive source for biopetroleum. While the chemistry of the hydrocarbons produced by the B race of B. braunii has been studied for many years, there is a deficiency of information concerning the molecular biology of this alga. In addition, there has been some discrepancy as to the phylogenetic placement of the Berkeley (or Showa) strain of the B race. To clarify its classification, we isolated the Berkeley strain nuclear SSU (18S) rRNA gene and β‐actin cDNA and used these sequences for phylogenetic analysis to determine that the Berkeley strain belongs to the Trebouxiophyceae class. This finding is in agreement with other B races of B. braunii, indicating the Berkeley strain is a true B race of B. braunii. To better understand molecular aspects of B. braunii, we obtained the Berkeley strain genome size as a first step in genome sequencing. Using flow cytometry, we determined the B. braunii Berkeley genome size to be 166.2 ± 2.2 Mb. We also estimated the GC content of the Berkeley strain as 54.4 ± 1.2% for expressed gene sequences.

MOLECULAR EVIDENCE CONFIRMS SISTER RELATIONSHIP OF ARDISSONEA, CLIMACOSPHENIA, AND TOXARIUM WITHIN THE BIPOLAR CENTRIC DIATOMS (BACILLARIOPHYTA, MEDIOPHYCEAE), AND CLADISTIC ANALYSES CONFIRM THAT EXTREMELY ELONGATED SHAPE HAS ARISEN TWICE IN THE DIATOMS(1).

With the use of a new kit from Qiagen to amplify total genome quantity, DNA was bulked up from two diatoms that are difficult to grow (Ardissonea and Climacosphenia), and the nuclear SSU rRNA gene was successfully amplified. Results of Bayesian analyses showed that these diatoms are sister to Toxarium and belong to the bi- and multipolar centric diatoms. The results indicate that extremely elongate shape has arisen at least twice in diatoms, in the true pennates, and in the bipolar centrics. The two lateral pattern centers of Ardissonea and Climacosphenia likely represent a modified annulus that subtends ribs internally as well as externally. Studies of sexual reproduction are needed to determine whether Ardissonea, Climacosphenia, and Toxarium achieve their elongate shape by similar means to each other and to true pennates, that is, by controlling the expansion of the auxospores by sequential addition of silicified bands (to form a properizonium or perizonium).

124 Evidence for Lateral Transfer of a IE Intron between Fungal and Red Algal Small Subunit Ribosomal RNA Genes

In a recent study of the North American biogeography of the red algae genus Hildenbrandia, the presence of group I introns were noted in the nuclear SSU rRNA gene of the marine species H. rubra (Hildenbrandiales). Group I introns in the nuclear encoded rRNAs have been previously reported in the Hildenbrandiales as well as the Bangiales. All reported introns within the red algae have been identified as belonging to the IC1 subclass and occur at two insertion sites in the nuclear small subunit rRNA (516 and 1506). However, an unclassified intron was discovered at position 989 in the nuclear SSU rRNA gene of a collection of H. rubra from British Columbia, Canada. We have determined that the intron is a member of the IE subclass and this is the first report of an IE intron and an intron in position 989 in the red algae. Phylogenetic analyses of the intron sequences reveal a close relationship between this group IE intron and similar ascomycete and basidiomycete fungal IE introns in the nuclear SSU rRNA genes at positions 989 and 1199. In addition, a common unique helix (structural signature) in the P13 domain of the Hildenbrandia intron and those of the fungi at the 989 and 1199 IE positions in the nuclear SSU rRNA gene also indicates a close relationship. Hence, this study provides evidence for a possible lateral transfer of the IE intron in position 989 between fungal and red algal nuclear SSU rRNA genes.